Cultivation bag assembly for cultivating microbes

ABSTRACT

A cultivation bag assembly for the cultivation of microbes, fungi and other organisms includes an inner bag for containing a food substrate and an organism to be cultivated. At least one wall of the inner bag is constructed of a layer of water-vapor permeable material. An outer bag of the assembly has first and second walls formed from gas impermeable layers. At least one of the first and second walls of the outer bag has a gas filter patch to allow the passage of oxygen and carbon dioxide gases into and out of the interior of the inner bag to facilitate incubation of the organism within the interior of the inner bag. The inner bag and outer bag are coupled together at a first end of each of the inner bag and outer bag.

FIELD OF THE INVENTION

The invention relates to cultivation bags for the cultivation ofmicrobes, fungi, and other organisms, their manufacture and use.

BACKGROUND

With higher demand for organic products, methods for increasing theproduction of crops without the use of chemical fertilizers, pesticidesand herbicides has been of particular interest and importance.Biopesticides used for agricultural purposes have been developed andcontinue to be developed to control pests and weeds, as well as act asfertilizers, thus increasing plant growth and crop yields without theneed for chemicals that can damage the environment and cause safetyand/or health concerns. These biological control agents are typically inthe form of natural microbes, fungi, or other organisms, and inparticular, fungi mycelium. These biological agents have a toxic effecton harmful insects and/or weeds, but do not negatively affect thegrowing crops, and/or may act beneficially as fertilizers that actuallypromote the growth of plants with which they are used.

The particular biological control agents used as pesticides act asparasitic microbes that attack the insects or undesirable plant mattercausing them to die. By spreading these microbes or biological agents inthe soil and fields, these materials can naturally destroy the damaginginsects and/or weeds, as well as act as fertilizing agents to facilitatecrop growth. Research and development is still underway in determiningand isolating effective microbes and biological agents out of the manythousands that exist for use as such pesticides. Therefore their use isonly expected to increase over time.

Of those parasitic microbes and organisms that are discovered andisolated for use as biopesticides, they must be produced in largequantities so that they can be used commercially for agriculturalpurposes. Once such system that is used by large industries involvesproduces the organisms in bulk. This involves the use of a large vesselcontaining a substrate. The substrate is a nutrient food source that amicrobe, fungi, or particular organism prefers and when introduced intothis substrate will tend to grow particularly rapidly on such substrate.In order to be used for cultivation the substrate must be sterilized toeliminate the growth of other undesirable microbes. To do this, thevessel with the substrate is typically heated to ≥121° C. for a certainlength of time. This is typically accomplished with the use of livesteam. The amount of time for sterilization depends on the amount ofsubstrate being sterilized. After sterilization, the vessel andsubstrate are cooled. The pure, isolated microbe or biological agent,such as fungi mycelium, which may be previously grown in petri dishes ortest tubes are then added to the substrate within the vessel. The vesselthen slowly rotates and mixes the materials until they are homogenouslymixed together.

Once the materials are mixed, the mixture is introduced into bags insuitable amounts and allowed to incubate at a desired temperature andhumidity to promote growth of the particular microbes or otherorganisms. Incubation is typically complete when the substrate iscompletely consumed. Incubation is ended by drying the bag contents,which typically occurs under vacuum. After drying, the resulting driedmicrobial material is crushed and reduced in size to desired particlessizes.

While the large bulk processing systems are useful in producing largequantities of microbial materials useful as biopesticides, they haveshortcomings. Such bulk systems are expensive and difficult to use,making them practical only for large producers. Furthermore, the bulkvessel is susceptible to contamination. If even one contamination sporeenters the bulk vessel after sterilization, such as during introductionof the desired microbe that is being cultivated, the whole batch may berendered unusable.

A more affordable method that can be used by smaller producers does notmake use of a bulk vessel. Instead smaller plastic bags filled withsubstrate are used that can withstand the temperatures of sterilization(i.e., ≥121° C.) for a sufficient amount of time to sterilize thesubstrate. The bags may have a small filter to allow steam used forsterilization to escape during the sterilization procedure so that thebag does not explode or burst as it is heated. If no filter is used, thebag may be positioned upright with one end of the bag being open butwith the opening restricted with either a foam or cotton wool materialto allow the escape of steam.

Once the sterilized substrate is cooled, pure microbes or organisms tobe cultivated are introduced into the bag. If a filtered bag is used,the open end is sealed, such as by welding. The bag and its contents arethen agitated to mix the materials and the bag is placed on a shelf forincubation.

After incubation, the bag is cut open and the contents are poured into asecond drying bag. Drying typically occurs with dry heating under avacuum within a vacuum chamber for increased efficiency. Drying in bagsis more suitable than in industrial drying, which involves highertemperatures, which can kill off the microbes that have been cultivated.The drying bag must be constructed to allow water vapor to readilyescape while retaining the bag contents within the bag. Thus, the dryingbag must have a large area of filter material to allow the water vaporto escape efficiently and quickly. The requirement of needing a seconddrying bag, however, adds additional costs and labor, as the ripemicrobes from the incubation bag must be transferred to the drying bagto facilitate drying.

To overcome the shortcomings of the above-described methods,improvements are needed.

BRIEF DESCRIPTION OF THE DRAWINGS

For a more complete understanding of the disclosure, reference is nowmade to the following descriptions taken in conjunction with theaccompanying figures, in which:

FIG. 1 is a perspective view of a cultivation bag assembly resting on asolid support surface and shown constructed in accordance withparticular embodiments of the invention;

FIG. 2 is an exploded perspective view of the cultivation bag assemblyof FIG. 1;

FIG. 3 is a cross-sectional view of cultivation bag assembly of FIG. 1taken along the lines 3-3;

FIG. 4 is a cross-sectional view of cultivation bag assembly of FIG. 1taken along the lines 4-4;

FIG. 5 is a perspective view of the cultivation bag assembly of FIG. 1shown with the forward end of an outer bag of the bag assembly foldedback to access the forward end of an inner bag of the bag assembly; and

FIG. 6 is a perspective view of the cultivation bag assembly of FIG. 1shown resting on a grate with the outer bag pulled away from the innerbag during a drying configuration.

DETAILED DESCRIPTION

The control of pests by interference with their ecological status isachieved by introducing a natural enemy or a pathogen into theenvironment. This is also called biocontrol. In simple terms, parasitesand agents that cause sickness to pests are introduced into the pests bynatural means and cause the pests to die. The present invention isrelated to an improved cultivation bag assembly that has many advantagesover the present art to reduce the steps in preparation and manufactureof these agents and to reduce risks of competing or contaminating matterintroduced into to the process.

The bag assemblies of the invention are useful in cultivation throughsolid state fermentation. Solid state fermentation (SSF) can be definedas the growth of microorganisms in a moist solid substance in theabsence of liquid water. The substance or substrate usually is composedof grain or grain waste, with addition of sugar, corn grits, rice wheatwaste, potato waste. etc., which mimic the natural environment which thebiological control agents grow in nature. SSF simulates the livingconditions of filamentous fungi and is the biotechnological process ofchoice. Under favorable conditions, spore germination takes placethrough the formation of tubes which grow and will be the base of futuremycelium. Through this process mycelium and spore production become thebiocontrol agent. This agent infects and kills the pests once it isintroduced into their environment.

As filamentous fungi grow hypha, which are the branching filaments thatmycelium of the fungi, penetrate into the solid matrix, becomingimpossible to separate substrate from mycelium. The biomass thus formedbecomes the product to be dried and reduced in size so that it can beeasily sprayed into fields or areas where the pests to be destroyedlive. Most of the time this will be on vegetation that are planted inagriculture. It is therefore an advantage to stimulate maximum growth offilamentous fungi spores by allowing wet substrate contained in a bagand providing a breathing means through a filter. The filter is designedto allow gas exchange between the filamentous fungi in the substrate andambient air by expelling CO₂ (carbon dioxide) generated in the bag andbreathing in O₂ (oxygen). The filter will also have small enough poresto obstruct contaminating living matter to enter the bag and thus toconsume and share the nutrients in the substrate. The amount of CO₂ inthe bag is critical to optimum growth of filamentous fungi.

By placing the incubating bag under controlled conditions oftemperature, ventilation, humidity and nutrients used, the substratemass is totally consumed by the cultivated biological control agent.When the substrate mass is fully consumed, the bag contents are dried.In the prior art methods, this is done by removing the mass from theincubation bag and introducing it into a drying bag. The presentinvention eliminates this stage, however, since the incubation anddrying bag are part of the same assembly.

In the drying process, drying air may be introduced into the environmentand the large physical area of an underlying permeable material of thedrying bag allows rapid dehydration of the biomass in the bag. Whendrying is completed, the drying bag is opened and the dried bio mass isreduced into particles that are further size reduced for packaging andmarketing.

Referring to FIG. 1, a cultivation or incubation bag assembly 10 isshown for cultivating and drying microbes, fungi and other organisms,such as fungi mycelium, as discussed. The various components of thecultivation bag assembly 10 and its construction can be seen morereadily in FIGS. 2 and 3.

As shown in FIGS. 2 and 3, the bag assembly 10 includes multiple layersof material that are joined together to form the bag assembly 10. Thebag assembly includes an inner drying bag 12 having flexible upper andlower walls 14, 16 joined around their perimeters along left- andright-side edges, such as at joint or seal 18 (FIG. 3) to define a baginterior 20 between the walls 14, 16. The front of the inner bag 12 mayinitially be open to allow for the introduction of a substrate materialand a microbe, fungi, or other organism to be cultivated.

In the embodiment shown, all or a major portion of the upper wall 14 isformed from a layer of gas and water-vapor permeable material. Thewater-vapor permeable material of the upper wall 14 is constructed toallow the passage or transmission of air and other gases, as well aswater vapor through the material of the upper layer 14. This may be anon-woven polyolefin fiber material, such as that sold under the nameTYVEK®, available from E. I. du Pont de Nemours and Company, Wilmington,Del. The polyolefin fiber material of the upper wall may bepolypropylene, polyethylene or high-density polyethylene (HDPE), or acombination of such materials. Examples of suitable commerciallyavailable products for the material of the upper wall 14 include, butare not limited to, TYVEK® 1025B, TYVEK® 1059B, and TYVEK® 1072Bmaterials. The permeable material of wall 14 may have a perm rating offrom 5 or greater to facilitate the passage of water vapor through thematerial. In particular embodiments, the water vapor transmissionthrough the layer 14 may range from at least, equal to, and/or betweenany two 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21,22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39,40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57,58, 59, and 60 perms or more, in certain instance. The permeablematerial of wall 14 may be suitable for filtering out particles having aparticle size of from 0.2 micron to 1 micron. Thus, the permeablematerial of wall 14 retains particles and powder of larger particlesizes within the inner drying bag 12.

It should be noted in the description, if a numerical value,concentration or range is presented, each numerical value should be readonce as modified by the term “about” (unless already expressly somodified), and then read again as not so modified unless otherwiseindicated in context. Also, in the description, it should be understoodthat an amount range listed or described as being useful, suitable, orthe like, is intended that any and every value within the range,including the end points, is to be considered as having been stated. Forexample, “a range of from 1 to 10” is to be read as indicating each andevery possible number along the continuum between about 1 and about 10.Thus, even if specific points within the range, or even no point withinthe range, are explicitly identified or referred to, it is to beunderstood that the inventor appreciates and understands that any andall points within the range are to be considered to have been specified,and that inventor possesses the entire range and all points within therange.

The lower wall 16 may be formed from the same or a similar permeablematerial as wall 14. If formed from the same material, the upper andlower walls 14, 16 may be formed by a single continuous sheet of suchmaterial by folding the sheet material along one edge so that the foldedlayers 14, 16 overlay one another. In such case, the seal, such as theseal 18, may be eliminated along such folded edge since the layers areeffectively joined together along the fold line.

In other embodiments, however, the lower wall 16 of inner bag 12 may beformed from a separate layer of flexible film that may be gasimpermeable so that water vapor and gases cannot pass or pass readilythrough the material of the film. The layer 16 may be formed from acontinuous thermoplastic film layer, such as the polyolefins ofpolypropylene, polyethylene or high-density polyethylene (HDPE). Incertain embodiments, the upper and lower walls 14, 16 may be configuredand joined together to provide gussets along the sides of the inner bag12 so that the inner bag 12 is a gusseted bag.

In many applications, the material of the upper and lower wall layers14, 16 may be of the same thermoplastic or polyolefin materials havingthe same or similar melting points, e.g., polypropylene, HDPE, etc., sothat they may be joined together through heat or ultrasonic welding sothat the materials are permanently or non-releasably fused together orotherwise strongly joined together so that they are not easilyseparated. In some instances, a different thermoplastic or polyolefinmaterial having different melting points may be used to join the layers14, 16 together. In other embodiments, other coupling means may be usedto join the layers 14, 16 at seal 18 along the left- and right-sideedges may be through the use of a strong adhesive layer positionedbetween the materials.

The bag assembly 10 further includes an outer incubation bag 22. Theouter incubation bag 22 is sized and configured to receive and enclosethe inner drying bag 12, including when the bag 12 is filled withsubstrate material and a microbe, fungi, or other organism to becultivated. The outer bag 22 has flexible upper and lower walls 24, 26joined around their perimeters along left- and right-side edges, such asat seam or seal 28 (FIG. 3) to define a bag interior 30 between thewalls 24, 26. The forward end of the outer bag 22 may initially be opento allow access to the forward end and opening 32 (FIG. 4) of the innerbag 12. The left- and right-side edges of the inner bag 12 are free fromor non-coupled to the outer bag 22.

The upper and lower walls 24, 26 of outer bag 22 may each be formed froma layer of flexible film that may be gas impermeable so that water vaporand gases cannot pass or pass readily through the material of the film.The layers 24, 26 may be transparent, opaque or non-transparent and mayeach be formed from a continuous thermoplastic film layer or sheet, suchas the polyolefins of polypropylene, polyethylene or high-densitypolyethylene (HDPE). In certain embodiments, the upper and lower walls24, 26 may be configured and joined together to provide gussets alongthe sides of the outer bag 22 so that the outer bag 22 is a gussetedbag.

In many applications, the material of the upper and lower wall layers24, 26 may be of the same thermoplastic or polyolefin materials havingthe same or similar melting points, e.g., polypropylene, HDPE, etc., sothat they may be joined together through heat or ultrasonic welding sothat the materials are permanently or non-releasably fused together orotherwise strongly joined together so that they are not easilyseparated. If formed from the same material, the upper and lower walls24, 26 may be formed by a single continuous sheet of such material byfolding the sheet material along one edge so that the folded layers 24,26 overlay one another. In such case, the seal, such as the seal 28, maybe eliminated along such folded edge since the layers are effectivelyjoined together along the fold line.

Referring to FIG. 4, the rearward end of the inner bag 12 is coupledtogether to the rearward end of the outer bag 22. This may be one ormore transverse seams or seals 34 that extend across all or a portion ofthe width of the each of the rearward ends of the inner and outer bags12, 22. As can be seen, the rearward end edges of the upper and lowerwalls 14, 16 of the inner bag 12 may be sandwiched between the rearwardend edges of the upper and lower walls 24, 26 of the outer bag 22. Thisseam or seal 34 also serves to couple the upper and lower layers 14, 16of the inner bag 12 to effectively close off the rearward end of theinner bag 12. Likewise, the rear end edges of the upper and lower walls24, 26 of the outer bag 22 are coupled along the rear end edges of theupper and lower walls 14, 16 along seam or seal 34 to effectively closeoff the rearward end of the outer bag 22.

In some embodiments, the materials of one or both of the upper and lowerwalls 14, 16 of the inner bag 12 and one or both of the upper and lowerwalls 24, 26 of the outer bag 22 may be formed from the samethermoplastic or polyolefin materials having the same or similar meltingpoints, e.g., polypropylene, HDPE, etc. This may allow the layers to bejoined together through heat or ultrasonic welding so that the materialsare permanently or non-releasably fused together or otherwise stronglyjoined together so that they are not easily separated. In someinstances, a different thermoplastic or polyolefin material havingdifferent melting points may be used to join one or both of the layers14, 16 of the inner bag 12 together and/or to one or both of the upperand lower layers 24, 26 of the outer bag 22 along seal 34. In otherembodiments, other coupling means may be used to join the layers 14, 16,24, 26 to one another along seal 34 along the rearward end edges may bethrough the use of a strong adhesive layers positioned between thematerials for permanent or non-releasable coupling.

In some instances, a different thermoplastic or polyolefin materialhaving different melting points or other properties may be used for oneor both of the upper and lower walls 14, 16 of the inner bag 12 fromthat of one or both of the upper and lower walls 24, 26 of the outer bag22. This difference in melting points or properties of the differentmaterials forming the layers 14, 16, 24, and 26 allows the materials tobe releasably coupled together. In certain embodiments, the differencein melting points between the different materials used to provide thereleasability may be from 10° C., 20° C., 30° C., 40° C., 50° C. ormore. Thus, for example, the layers 14, 16 of the inner bag 12 may beconstructed of polypropylene, which may have a melting point of from130° C. to 171° C. The layers 24, 26 of the outer bag 22, however, maybe formed from HDPE, which may have a melting point of from 115° C. to135° C. Because of the different materials used and their differentmelting points and/or properties, the materials are not as strongly heldtogether along seal or seam 34 as they would be if they were formed fromthe same materials. This low bonding strength may aid or facilitate thereleasability of the layers 14, 16, 24, 26 from one another along seam34, as will be discussed later on. In other instances, the use of areleasable adhesive may be used to join the layers 14, 16, 24, 26 alongseam 34. This may be a continuous layer of adhesive that effectivelyseals and releasably joins the layers together.

Referring to FIG. 2, an opening or slot 36 is formed in the upper wall24 of the outer incubation bag 22 at a position so that the opening 36directly overlays the permeable layer of the upper wall 14 of inner bag12 when the inner drying bag 12 is positioned within the outercultivation bag 22. The opening or slot 36 is shown as being anelongated slot that extends along a major portion of the length of theupper wall 24. The opening 36 is smaller in area than the permeableupper wall 14 so that only a portion of the upper wall 14 underlies theopening 36.

A gas filter patch 38 is coupled to the upper wall 24 and is used tocover the opening 36. The gas filter patch 38 is an incubation filterthat allows the optimum gas exchange into the interior 30 of the outerincubation bag 22 and into the interior 20 of the inner drying bag 12.This involves the passage of air and/or oxygen and carbon dioxide gasesto and from ambient air, as well as some water vapor, to facilitatecultivation. The gas filter patch 38 may a non-woven polyolefin fibermaterial. There are many commercially available materials useful for thefilter patch 38, which may be formed from non-woven polypropylene orHDPE. The filter patch material will typically have smaller openingsthan that used for the permeable layer of upper wall 14 of inner dryingbag 12. In particular applications, the filter patch 38 may be suitablefor filtering out particles having a particle size of from 0.01 micronto 0.2 micron or greater. The filter patch 38 and the correspondingopening 36, in conjunction with the porosity of the filter patch 38, maybe sized and configured to allow the selected passage of air, oxygen andcarbon dioxide, and some water vapor, for the particular organism beingcultivated. Thus, the size and configuration of the filter patch 38 andopening 36 may vary depending on the purpose, use, and environmentalconditions (i.e., the type of organism being cultivated and itscultivating environment) of the bag assembly 10.

The side edges of the filter patch 38 are coupled or joined to the upperlayer 24 of the outer bag 22, around the entire perimeter of the opening36 so that the opening 36 is completely covered by the filter patch 38.In many applications, the material of the upper film layer 24 and thefilter patch layer 38 may be of the same thermoplastic or polyolefinmaterial having the same or similar melting points, e.g., polypropyleneor HDPE, so that they may be joined together through heat or ultrasonicwelding so that the materials are permanently or non-releasably fusedtogether or otherwise strongly joined together so that they are noteasily separated. In other embodiments, other coupling means may be usedto join the filter patch 38 to the film layer 24, such as through theuse of a strong adhesive layer positioned between the materials.

In forming the bag assembly 10, the layers 14, 16 of the inner bag 12are coupled together along their left- and right-side edges along thelength of the layers 14, 16. This may be done by heat or ultrasonicwelding, as previously discussed, so that the materials are permanentlyor non-releasably fused together or otherwise strongly joined together.Alternatively, a strong adhesive may be used to couple the layers 14,16. In manufacturing, long, continuous lengths of the sheet materialforming the layers 14, 16 may be coupled together in this fashion withthe seams or seals 18 extending along the continuous length of thelayers. The coupled layers may then be cut transversely to the desiredlength to form several of the individual inner bags 12. At this stagethe forward and rearward ends of the inner bag 12 remain open.

Likewise, the layers 24, 26 of the outer bag 22 are coupled togetheralong their left- and right-side edges along the length of the layers24, 26. This may also be done by heat or ultrasonic welding, aspreviously discussed, so that the materials are permanently ornon-releasably fused together or otherwise strongly joined together.Alternatively, a strong adhesive may be used to couple the layers 24,26. In manufacturing, long, continuous lengths of the sheet materialforming the layers 24, 26 may be coupled together in this fashion withthe seams or seals 28 extending along the continuous length of the layermaterials. The coupled layers may then be cut transversely to thedesired length to form several of the individual outer bags 22. At thisstage the forward and rearward ends of the outer bag 22 also remainopen.

The thus formed inner bag 12 is positioned within the interior of theouter bag 22. The inner bag 12 will typically have a smaller width andlength than the outer bag 22 to facilitate fitting and positioning ofthe inner bag 12 within the outer bag 22. With the inner bag 12positioned within the interior of the outer bag, the inner and outerbags 12, 22 are coupled together at their rearward ends along transverseseal or seam 34, as previously discussed, so that each of the rearwardends of the inner and outer bags 12, 22 are effectively closed and theinner bag 12 is coupled to the outer bag 22 along the seam 34. In thisconfiguration, each of the upper and lower walls 14, 16 and 24, 26 ofthe inner and outer bags 12, 24 will project forward from the seam orseal 34 in the same direction, with the left- and right-side edges ofeach bag 12, 22 being adjacent to and extending generally parallel toone another, with the inner bag 12 being contained within the outer bag22.

As shown in FIG. 5, the end portions of layers 24, 26 forming the openforward end of the outer bag 22 are initially folded back uponthemselves so that the forward end of the inner bag 12 projects out adistance beyond the forward opening of the outer bag 22. In thisconfiguration, the forward end of the inner bag 12 is open with thefront-end edges of the upper and lower walls 14, 16 not being joinedtogether to define the opening 32 of the inner bag 12. This allows forthe introduction of a food substrate, along with microbes, fungi orother organisms, such as fungi mycelium, to be cultivated, such as shownat 44 of FIGS. 3 and 4, into the inner bag interior 20. The foodsubstrate is typically an organic material that provides nutrients andmoisture for the cultivation of the desired microbe or organisms. Thesubstrate may be wet or dry. One suitable type of substrate useful forthe cultivation of fungi mycelium is wet rice. Other substrate materialsmay also be used, however, and may vary based upon the organism beingcultivated.

The bag assembly 10, including any substrate, may be sterilized orfabricated under sterile conditions to prevent contamination of thecultivated organisms. If the materials can withstand heat sterilization,heat sterilization may be used. Heat sterilization may be used byheating the bag assembly 10 and/or its components, with or without anysubstrate, to a temperature of from 121° C. or higher to effectivelykill any unwanted bacteria, viruses or other undesirable microorganisms.

Because components of the bag assembly 10 may be constructed ofdifferent thermoplastic materials, such as HDPE, which may have a lowermelting point that would degrade the material if such high temperatureswere used, other non-heat sterilization techniques may need to be used.This may include radiation, such as gamma radiation, or chemicalsterilization. In such instances, the bag assembly 10, with or withoutany substrate, is subjected to radiation or sterilization chemicals toeffectively sterilize the bag assembly and any substrate containedtherein.

Once the bag assembly 10 and substrate are sterilized, puremicroorganisms, fungi or other organisms to be cultivated are introducedinto the interior 20 of the inner bag 12 through opening 32. This may bedone in sterile or clean room conditions to avoid any contamination ofthe materials.

With the food substrate and microbe, fungi or organisms to be cultivatedlocated within the interior 20 of the inner bag 12, the layers 14, 16can be joined or coupled together at their forward end edges acrosstheir widths, such as along dashed line 46, so that the opening 32 issealed or closed and these materials are enclosed within the interior 20of the inner bag 12. This may be through heat or ultrasonic welding sothat the materials are permanently or non-releasably fused together orotherwise strongly joined together so that they are not easilyseparated. In other embodiments, other coupling means may be used tojoin the end edges of the layers 14, 16 together, such as through theuse of a strong adhesive layer positioned between the materials.

The forward end edges of the layers 24, 26 of the outer bag are thenunfolded, such as shown by the arrows 48, so that the layers 24, 26cover the now closed end of the inner bag 12. The unfolded ends oflayers 24, 26 can then be joined or coupled together at their forwardends across their widths, so that the forward opening of the outer bag22 is closed and the inner bag 12 is contained enclosed within theinterior 30 of the outer bag 22 in a cultivation configuration of thebag assembly 10. The joining of the layers 24, 26 at their forward endsmay be through heat or ultrasonic welding so that the materials arepermanently or non-releasably fused together or otherwise stronglyjoined together so that they are not easily separated. In otherembodiments, other coupling means may be used to join the forward endedges of the layers 24, 26 together, such as through the use of a strongadhesive layer positioned between the materials.

After the bag assembly 10 is filled and sealed, the bag assembly 10 andits contents may then be agitated to thoroughly mix the contents withinthe bag assembly. With the cultivation bag assembly 10 sealed and thecontents mixed, the bag assembly 10 and its contents are placed in anenvironment (e.g., temperature, humidity, light, etc.) suitable forgrowing and cultivation. As shown in FIG. 1, the bag assembly 10 may beplaced upon a solid shelf or support surface during cultivation, such asthe support surface 40. The support surface 40 may be solid plastic,stainless steel, wood, concrete, etc. The bag assembly 10 may also beused over a non-solid or vented surface during cultivation, such as thegrating 42 of FIG. 6. This may facilitate drying during the drying step,which is discussed later. During cultivation, the gas filter patch 38positioned over the opening 36 of the upper wall 24 of outer bag 22 isexposed and allows the optimum gas exchange into the interior 30 of theouter bag 22 and through the permeable upper wall 14 into the interior20 of the inner bag 12 where the substrate and cultivated organisms arelocated. This involves the passage of air and/or oxygen and carbondioxide gases to and from ambient air, as well as some water vapor, tofacilitate cultivation.

When the cultivated organisms are ripe, which is typically after all ofthe substrate has been consumed by the cultivated organism, the bagassembly contents are dried. As discussed earlier, in prior art methods,this would typically occur by emptying the contents of the cultivationbag into a separate drying bag. With the bag assembly 10 of theinvention, however, this is not needed as both cultivation and dryingcan be accomplished using the same bag assembly.

To dry the bag assembly contents, the outer bag 22 is opened at theforward end and pulled back away from the inner bag 12 so that the innerbag 12 is exposed in a drying configuration of the bag assembly 10, asshown in FIG. 6. In this drying configuration, the upper and lower walls24, 26 of the outer bag 22 will be folded back over the seal or seam 34that couples the inner bag 12 and outer bag 22 together so that theinner bag 12 is no longer contained within the interior of the outer bag22 and the outer bag 22 is now folded inside out. Water vapor passesthrough the water-vapor permeable upper layer 14 of the inner bag 12, tofacilitate drying. In many applications, the drying may be conductedwith heating and/or under vacuum within a vacuum chamber to speed updrying. In applications, where the bag assembly 12 is used on a grating,such as grating 42 of FIG. 6, the lower wall 16 of the inner bag mayalso be formed from a permeable material, such as the permeable materialof the upper wall 14, to further facilitate drying.

During drying, the outer bag 22 may remain coupled at its rearward endalong seam 34 to the rearward end of the inner bag 12 but be moved to aposition so that the permeable upper wall 14 of the inner bag 12 remainsexposed to facilitate drying. In other embodiments, however, the outerbag 22 may be removed completely from the inner bag 12. In suchinstances, the materials of the outer bag 22 may be coupled to thematerials of the inner bag 12 with a low bond strength to facilitateremoval of the outer bag 22. This may be accomplished by using differentmaterials having different melting points for the upper and lower walls14, 16 of the inner bag 12 and the upper and lower walls 24, 26 of theouter bag 22, as discussed previously. The difference in melting pointsor properties of the different materials forming the layers 14, 16 and24, 26 creates a low bonding strength that allows the materials to bereleasably coupled together. In other instances, the use of a releasableadhesive may be used to join the layers 14, 16 and 24, 26 along seam 34.This may be a continuous layer of adhesive that effectively seals andreleasably joins the layers together. Alternatively, the outer bag 22may be cut away from the inner bag 12 if it is no longer needed.

After drying, the inner drying bag 12 of the bag assembly 10 and itscontents may be agitated to break the materials located within intosmaller particles. The inner bag 12, with or without the outer bag 22,and its contents may then be packaged, shipped and/or stored for lateruse as biopesticide or biological agent that can be spread onagricultural plots for growing crops for controlling pests, weeds or foruse as a fertilizer, as described earlier. The inner bag 12 and bagassembly 10 is made of durable materials so that it can be used forshipping and storage without having to unseal and remove the contentsand placing them in a different bag prior to use.

While the invention has been shown in only some of its forms, it shouldbe apparent to those skilled in the art that it is not so limited but issusceptible to various changes and modifications without departing fromthe scope of the invention. Accordingly, it is appropriate that theappended claims be construed broadly and in a manner consistent with thescope of the invention.

I claim:
 1. A cultivation bag assembly for the cultivation of microbes,fungi and other organisms comprising: an inner bag having first andsecond walls that are joined together along side edges to define aninner bag interior for containing a food substrate and an organism to becultivated, at least one of the first and second walls being constructedof a layer of water-vapor permeable material to allow the passage ofwater vapor therethrough; and an outer bag having first and second wallsjoined along side edges to define an outer bag interior, the first andsecond walls of the outer bag being formed from gas impermeable layers,at least one of the first and second walls of the outer bag having anopening with a gas filter patch covering the opening to allow thepassage of oxygen and carbon dioxide gases through the gas filter patchto and from ambient air and into and out of the interior of the innerbag to facilitate incubation of the organism within the interior of theinner bag, and wherein the inner bag and outer bag are coupled togetherat a first end of each of the inner bag and outer bag, with the innerbag being contained within the interior of the outer bag in acultivation configuration.
 2. The bag assembly of claim 1, wherein: bothfirst and second walls of the inner bag are each constructed of a layerof water-vapor permeable material.
 3. The bag assembly of claim 2,wherein: the layer of water-vapor permeable material is constructed froma non-woven polyolefin fiber material.
 4. The bag assembly of claim 1,wherein: the inner bag and outer bag are coupled together at the firstend to close the first end of each of the inner bag and outer bag. 5.The bag assembly of claim 1, wherein: the outer bag and the inner bagare formed from at least one of polypropylene, polyethylene, and highdensity polyethelene (HDPE).
 6. The bag assembly of claim 1, wherein:the side edges of the inner bag are non-coupled to the outer bag.
 7. Thebag assembly of claim 1, wherein: the bag assembly is a sterilized bagassembly.
 8. The bag assembly of claim 1, wherein: the water-vaporpermeable material has a perm rating of from 5 or greater.
 9. The bagassembly of claim 1, wherein: a second end of each of the inner bag andthe outer bag are non-coupled together.
 10. The bag assembly of claim 9,wherein: the interior of the inner bag contains at least one ofmicrobes, fungi and other organisms to be cultivated together with afood substrate, with the first and second walls of the inner bag beingjoined together at the second end of the inner bag to enclose said atleast one of microbes, fungi and other organisms and food substratewithin the interior of the inner bag; and wherein the first and secondwalls of the outer bag are joined together at the second end of theouter bag to enclose the inner bag within the interior of the outer bag.11. A method of forming a cultivation bag assembly for the cultivationof microbes, fungi and other organisms comprising: forming an inner baghaving first and second walls that are joined together along side edgesto define an inner bag interior for containing a food substrate and anorganism to be cultivated, at least one of the first and second wallsbeing constructed of a layer of water-vapor permeable material to allowthe passage of water vapor therethrough; and forming an outer bag havingfirst and second walls joined along side edges to define an outer baginterior, the inner bag being contained within the interior of the outerbag, the first and second walls of the outer bag being formed from gasimpermeable layers, at least one of the first and second walls of theouter bag having an opening with a gas filter patch covering the openingto allow the passage of oxygen and carbon dioxide gases through the gasfilter patch to and from ambient air and into and out of the interior ofthe inner bag to facilitate incubation of the organism within theinterior of the inner bag; and coupling the inner bag and outer bagtogether at a first end of each of the inner bag and outer bag to closethe first end of each of the inner bag and outer bag.
 12. The method ofclaim 11, wherein: each of the inner bag and outer bag has an opensecond end; and further comprising: introducing at least one ofmicrobes, fungi and other organisms to be cultivated together with afood substrate into the interior of the inner bag through the opensecond end of the inner bag; joining the first and second walls of theinner bag together at the second end of the inner bag to enclose said atleast one of microbes, fungi and other organisms and food substratewithin the interior of the inner bag; and joining the first and secondwalls of the outer bag together at the second end of the outer bag toenclose the inner bag within the interior of the outer bag.
 13. Themethod of claim 12, wherein: the end portions of the first and secondwalls of the outer bag at the second end of the outer bag are initiallyfolded back upon themselves so that the first and second walls of theinner bag at the second end of the inner bag project beyond the secondend of the outer bag; and further comprising: joining the projectingfirst and second walls of the inner bag together at the second end ofthe inner bag to close the second end of the inner bag.
 14. The methodof claim 13, further comprising: unfolding the end portions of the firstand second walls of the outer bag at the second end of the outer bag;and joining the unfolded end portions of the first and second walls ofthe outer bag together at the second end of the outer bag so that theinner bag is enclosed within the interior of the outer bag.
 15. Themethod of claim 11, wherein: both first and second walls of the innerbag are each constructed of a layer of water-vapor permeable material.16. The method of claim 11, wherein: the outer bag and the inner bag areformed from at least one of polypropylene, polyethylene, and highdensity polyethelene (HDPE).
 17. The method of claim 11, wherein: theside edges of the inner bag are non-coupled to the outer bag.
 18. Themethod of claim 11, wherein: the bag assembly is a sterilized bagassembly.
 19. The method of claim 11, wherein: the water-vapor permeablematerial has a perm rating of from 5 or greater.
 20. The method of claim11, wherein: a second end of each of the inner bag and the outer bag arenon-coupled together.